Monoclonal Antibodies and the Rise of Precision Immunopsychiatry

en Monoclonal Antibodies and the Rise of Precision Immunopsychiatry 94 95 https://www.ajmb.org/En/Article.aspx?id=70644 https://www.ajmb.org/PDF/En/FullText/70644.pdf ShahinAkhondzadeh739

en Luteolin-Treated 4T1 Cell-Derived Exosomes as Novel Antiproliferative Agents In Vitro and In Vivo Background: Breast cancer is the most widespread malignancy among women worldwide. Luteolin, a flavonoid, has demonstrated anti-cancer effects by triggering apoptosis in tumor cells. Exosomes are gaining much attention for cancer therapeutic approaches due to multitude of beneficial effects. This study is aimed to investigate the possible potential of exosomes derived from luteolin-treated 4T1 cells to ameliorate tumor in comparison to luteolin treatment only. Methods: In this study, 4T1 cell culture was exposed to luteolin. Following exosome extraction, they were characterized using field emission scanning electron microscopy, dynamic light scattering and western blot analysis. MTT assay was performed in order to evaluate cell viability after exposure to different concentrations of luteolin and exosomes. An in vivo breast cancer model was induced via subcutaneous injection of 4T1 cells to the BALB/C mice. After 14 days, tumor volume was measured, and expression of RhoA and ERK mRNAs were quantified by Real Time PCR. Results: The MTT assay demonstrated that exosomes from luteolin-treated 4T1 cells at a concentration of 320 μg/μl reduced cell viability by approximately 70% in a dose-dependent manner. Tumor volume in the exosome-treated group decreased by 57% relative to the tumor group, while the luteolin-treated group demonstrated a 39% reduction. Furthermore, RhoA gene expression was substantially downregulated in the exosome-treated group, and exosomes were more effective than luteolin in reducing ERK gene expression. Conclusion: Exosomes derived from luteolin-treated 4T1 cells effectively suppress breast cancer cell growth by reducing 4T1 cell viability and by decreasing tumor volume and downregulating tumor-associated genes RhoA and ERK. These results propose a novel therapeutic strategy for breast cancer, highlighting the promising potential of exosomes as an efficient drug delivery system. Breast neoplasms, Cell survival, Exosomes, Luteolin, Tumor burden 96 103 https://www.ajmb.org/En/Article.aspx?id=70645 https://www.ajmb.org/PDF/En/FullText/70645.pdf PardisHeidariFaculty of Veterinary Sciences, SR.C., Islamic Azad University, Tehran, Iran92480RaziehHosseini92481Nika Nasihatgar Faculty of Veterinary Sciences, SR.C., Islamic Azad University, Tehran, Iran92482

en Investigation of the Molecular Signature of Senescence in Mesenchymal Stem Cells Background: Mesenchymal Stem Cells (MSCs) play a pivotal role in regenerative medicine due to their multipotency and immunomodulatory properties. However, during in vitro expansion, MSCs undergo senescence, characterized by a decline in proliferation, impairment of differentiation potential, and altered secretory profiles, which limits their therapeutic efficacy. This study aimed to identify novel molecular regulators and net-work-level interactions underlying MSC senescence through microarray analysis of the GSE7888 dataset, comparing early and senescent MSCs. Methods: A total of 4597 Differentially Expressed Genes (DEGs) were identified be-tween early (passages 4-5) and senescent (passages 22-28) MSCs, with 2219 upregulated and 2379 downregulated. Key regulators such as CDKN1A (p21), CDK4, and CDK6 were implicated in cell cycle arrest and the progression of senescence. Pathway analysis highlighted the mTOR, FoxO, and p53 signaling pathways as key regulators of stress responses, metabolism, and aging. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis highlighted cellular processes, including protein transport and intracellular signaling, while protein-protein interaction networks identified high-connectivity nodes such as TP53, FOXO3, and MDM2. Senescent MSCs displayed phenotypic changes, including altered morphology and the emergence of the Senescence-Associated Secretory Phenotype (SASP), which impaired re-generative potential. Results: The findings suggest that targeting the mTOR, FoxO, and p53 pathways could delay senescence and enhance MSC therapeutic potential. Interventions such as rapamycin and FoxO3 activators show promise in reversing senescence. Conclusion: Future research should explore small molecules and gene-based therapies targeting senescence pathways to improve MSC-based regenerative strategies. These results provide a foundation for developing innovative approaches to optimize MSC ap-plications in clinical therapies. Gene ontology, Mesenchymal stem cells, Regenerative medicine, Senescence-associated secretory phenotype 104 112 https://www.ajmb.org/En/Article.aspx?id=70646 https://www.ajmb.org/PDF/En/FullText/70646.pdf Forough-AzamSayahpour Medical College of Wisconsin, Center of Cancer Discovery, Milwaukee, Milwaukee, Wisconsin, United States of America92483MarjanNejatiDepartment of Cell and Molecular Biology, School of Biology, College of Sciences, University of Tehran, Tehran, Iran92484MahyaRouhollahi-Masoumi Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biolo-gy and Technology, ACECR, Tehran, Iran92485MohammadrezaBaghaban Eslaminejad1313SaraTaleahmad92487

en Potential Targets in Innate Immunity Receptors for Gastric Cancer: Insights from Virtual Screening in TCM and In Vitro Assay Background: Gastric Cancer (GC) poses a substantial global health threat, ranking as the second leading cause of cancer-related mortality among gastrointestinal malignancies. This investigation explores the potential therapeutic implications of plant extracts on gastric cancer, with a specific focus on their effects on the innate immune system. Methods: A comprehensive analysis was conducted using 200 Sequence Read Runs (SRRs) thigh samples associated with gastrointestinal cancer tissue, juxtaposed with pathologically confirmed healthy tissues serving as controls. Differential Gene Expression (DGE) testing, encompassing the examination of 28,000 genes, including 95 pivotal genes associated with the innate immune system, was conducted. Findings elucidate alterations in the expression of key pattern recognition receptors, such as TLR2 and TLR4, as well as pivotal molecules within their signaling pathways. In pursuit of potential antagonists for these receptors, virtual screening on the Maestro docking platform in the Schrödinger 2022 package was conducted, evaluating 220,000 diverse tautomer’s of plant active substances. Selected candidates, exhibiting superior docking scores across four additional platforms, were subjected to further scrutiny. Results: MMT results showed that nettle extract showed significant cytotoxic effects within 12 hr compared to the control (no treatment) sample, resulting in a 34.7% reduction in AGS cancer cell viability. The flow cytometry test showed that the control group had 71%, and groups treated with nettle extract for two and 12 hr had 65.3 and 67.18% viable cells, respectively. These differences were not statistically significant, indicating that nettle extract selectively preserves healthy living cells. Conclusion: Cytotoxicity tests and cell cycle assessments confirmed the ability of nettle extract to reduce the survival of GC cells. This property makes nettle a promising candidate for drug development in this direction. Cell cycle, Drug development, Gene expression, Immune system, Plant extracts, Signal transduction, Toll-Like Receptor 2, Toll-Like Receptor 4 113 120 https://www.ajmb.org/En/Article.aspx?id=70647 https://www.ajmb.org/PDF/En/FullText/70647.pdf AbbasGanjaliUniversity of Zabol, Zabol, Iran92488BarataliFakheriDepartment of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Zabol, Zabol, Iran92489AbbasBahari92490LeilaFahmideh Department of Plant Breeding and Biotechnology, Gorgan University of Agriculture Sciences and Natural Resources, Gorgan, Iran92491RezaValadanDepartment of Immunology, Molecular and Cell Biology Research Centre, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran92492MehdiTavakolizadeh Department of Pharmacognosy, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran92493

en Therapeutic Potential of Alternating Magnetic Fields for Normalizing Blood Parameters and Restoring Renal, and Cardiac Function in Diabetic Mice Background: In recent years, the number of adults aged 20-79 years living with diabetes has increased more than threefold. Currently, the treatment of diabetes typically involves the long-term use of chemical and herbal drugs. However, prolonged use of chemical drugs may lead to side effects that can be detrimental to health. Therefore, this study aims to normalize blood glucose levels and restore kidney and heart cells. Methods: The research was conducted using diabetic mice as experimental subjects. The treatment involved exposure to an alternating Magnetic Field with Magnetic Flux Densities of 0.3 and 0.6 mT for 20 min/day over five consecutive days. The frequencies of the applied Magnetic Fields were 50, 100, 150, and 200 Hz. Results: The results showed that the greatest reduction in blood glucose levels (92.11%) was observed at a frequency of 100 Hz and a Magnetic Flux Density of 0.6 mT. Meanwhile, the highest increase in hemoglobin levels (81.11%) occurred at a frequency of 150 Hz and a Magnetic Flux Density of 0.3 mT. Other parameters that experienced non-linear changes included cholesterol levels, blood viscosity, and erythrocytes count, glomerulus and kidney cell density, and heart cell density. Conclusion: The optimal effects of magnetic field exposure do not always occur at the same frequency or Magnetic Flux Density. Blood, Blood glucose, Diabetes mellitus, Heart, Kidney 121 130 https://www.ajmb.org/En/Article.aspx?id=70648 https://www.ajmb.org/PDF/En/FullText/70648.pdf MokhamadTirono81834FaridSamsu HanantoDepartment of Physics, Faculty of Science and Technology, State Islamic University of Maulana Malik Ibrahim of Malang, Malang City, East Java, Indonesia81835

en Evaluation of the Anti-melanogenic Effect of Aqueous and Hydroalcoholic Extracts of Nasturtium officinale on the B16F10 Cell Line Background: This project aimed to evaluate the anti-melanogenic characteristics of Nasturtium officinale (N. officinale) by assessing the impact of both aqueous and hydroalcoholic extracts on the inhibition of cellular and mushroom tyrosinase enzymes, as well as the suppression of the melanin synthesis in B16F10 melanoma cells. Methods: The aerial components of N. officinale were subjected to extraction using distilled water: ethanol (7:3) through the maceration technique. The extract’s phenolic compounds were quantified employing the Folin-Ciocalteu method. The evaluation of the safety profile of the extracts on B16F10 cells was done by the MTT assay. Subsequently, the melanin concentration in B16F10 cells, alongside the inhibitory effects on both mushroom and cellular tyrosinase, was assessed following treatment with the aforementioned extracts. Results: The aqueous and hydroalcoholic extracts exhibited no significant toxicity on B16F10 when compared to Phosphate-Buffered Saline (PBS). Additionally, there was no notable difference in the cytotoxic effects of extracts on the B16F10 cell line. Both extracts resulted in inhibition of cellular and mushroom tyrosinase, along with a decrease in melanin levels in B16F10 in a concentration-dependent manner. Ultimately, the total phenolic content in the aqueous and hydroalcoholic extracts was found to be approximately 14 and 30 mg/g of gallic acid, respectively. Conclusion: This in vitro investigation offers evidence supporting the skin brightening properties of N. officinale as an anti-melanogenic agent. Given its safety profile and absence of toxic effects on melanoma cells, it may be incorporated into the formulation of skin-brightening products following preclinical tests. Melanin, Melanoma cell line, Nasturtium officinale, Tyrosinase 131 136 https://www.ajmb.org/En/Article.aspx?id=70649 https://www.ajmb.org/PDF/En/FullText/70649.pdf MohaddeseHeydari Department of Pharmaceutical Biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran92496MasoudSadeghi-Dinani Department of Pharmacognosy, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran92497FatemehShafiee92265

en Establishing an Optimized Caco-2/THP-1 Co-Culture Model to Efficiently Simulate Inflammatory Bowel Disease In Vitro Background: Inflammatory Bowel Disease (IBD) is a complex disorder for which the mechanisms and targeted therapies remain unclear. Several in vitro models, including organoids, cytokine-stimulated Caco-2 monolayers, and co-culture systems, have been developed to study IBD pathogenesis and potential treatments. Meanwhile, the Caco-2/THP-1 co-culture is a practical model representing the interaction of intestinal epithelial and immune cells. However, multiple factors, such as culture duration and exposure time to inflammatory agents, significantly affect model outcomes. Developing an optimized co-culture that better mimics intestinal inflammation can introduce a valuable method for future studies. This study aimed to optimize a Caco-2/THP-1 co-culture model, focusing on culture timing and treatment conditions. Methods: THP-1 monocytes were differentiated into macrophage-like cells (M0) with phorbol 12-myristate 13-acetate (PMA, 50 ng/ml, 48 hr). M0 cells were treated with different Lipopolysaccharide (LPS) concentrations for 6 or 24 hr to determine the optimal inflammatory dose. Inflammatory macrophages (M1) were co-cultured with differentiated or undifferentiated Caco-2 monolayers. Expression of IL-6, IL-8, and TNF-α was measured by qRT-PCR, M1 macrophage markers (CD86/HLA-DR) by flow cytometry, and nitric oxide by the Griess assay. Results: Stimulation with 100 ng/ml LPS for 6 hr increased M1 (CD86⁺/HLA-DR⁺) macrophages to 58.9% and induced maximal nitric oxide production (179.3 µM). Co-culture with these M1 cells enhanced IL-8 and modestly increased IL-6 expression in differentiated Caco-2 cells compared with other groups. Conclusion: The differentiated Caco-2/THP-1 co-culture efficiently mimics intestinal inflammation observed in IBD and provides an optimized in vitro model for further investigations. Biological models, Caco-2 cells, Inflammatory bowel disease, Lipopeptides, Macrophage activation 137 146 https://www.ajmb.org/En/Article.aspx?id=70650 https://www.ajmb.org/PDF/En/FullText/70650.pdf GildaParsamanesh Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran92499KavehBaghaeiOlivia Newton-John cancer Research Institute and School of cancer Medicine, La Trobe University, Heidelberg VIC, Australia92500NimaRezaeiNetwork of Immunity in Infection, Malignancy and Autoimmunity (NIIMA), Universal Scientific Education and Research Network (USERN), Tehran, Iran186

en Bioactive Silver Nanoparticles Coated with Curcumin Inhibit Angiogenesis through MMP-9 and Cox-2 Down-Regulation Background: Inhibition of angiogenesis is an attractive approach in cancer therapy. Both curcumin and silver nanoparticles (AgNPs) have demonstrated anti-angiogenic properties; however, the the poor water solubility of Curcumin and the side effects of AgNPs adversely affect their activity. Methods: In this study, AgNPs coated with curcumin (Cur-AgNPs), was used to improve aqueous-phase solubility of curcumin and decrease the side effects of AgNPs. After-wards, treatment with curcumin enhanced the anti-angiogenic activity of Cur-AgNPs. The nanoparticles were synthesized as both reducing and stabilizing agents. Evaluation of anti-angiogenesis was assessed in vitro using Human Umbilical Vein Endothelial Cells (HUVECs) and in vivo through the Chorioallantoic Membrane (CAM) assay. Da-ta were analyzed by one-way ANOVA with Tukey’s multiple comparison test. Results: Synthesized Cur-AgNPs have an average diameter of 39 nm, with spherical shapes and an absorbance peak at 450 nm in the UV-visible spectrum. Cur-AgNPs showed a negative zeta potential. EDAX and FTIR confirmed the conjugation of cur-cumin with AgNPs. In vitro anti-angiogenesis assays demonstrated that Cur-AgNPs re-duced the viability of HUVECs, an Inhibitory Concentration (IC50) value of 13 µg/ml. DAPI and acridine orange/propidium iodide staining revealed a significant increase in apoptotic cells following treatment with Cur-AgNPs. The expression of Matrix Metallo-proteinase 9 (MMP-9) and Cyclooxygenase-2 (COX-2) was also inhibited in treated cells. In vivo anti-angiogenesis assays using the CAM model showed significant de-crease in the number, length and hemoglobin content of CAM blood vessels. Conclusion: Curcumin conjugated with AgNPs may represent a promising strategy to enhance the therapeutic potential of both AgNPs and curcumin. However, further in-vestigations, particularly regarding safety and biocompatibility of Cur-AgNPs, is needed in this field. Angiogenesis, Curcumin, Nanoparticles, Silver 147 156 https://www.ajmb.org/En/Article.aspx?id=70651 https://www.ajmb.org/PDF/En/FullText/70651.pdf TayebeRamezani Farzin 92501MohammadNabiuni Department of Biology, Kharazmi University, Tehran, Iran92502

en Identification of a Novel Nucleic Acid Target for the Rapid and Specific Detection of Mycobacterium Simiae Using Comparative Genomic Analysis Background: Mycobacterium simiae (M. simiae) is a non-tuberculous mycobacterium (NTM) that closely resembles Mycobacterium tuberculosis (M. tuberculosis) in clinical and biochemical characteristics, notably its niacin-positive phenotype. This similarity frequently leads to misdiagnosis and inappropriate treatment with first-line anti-tuberculosis drugs, to which M. simiae is frequently resistant. Current diagnostic methods are expensive or need complex equipment, highlighting the urgent need for a rapid, specific, and accessible molecular target to identify M. simiae accurately. Methods: In this study, a modified genome comparison method was applied to the complete reference genome of M. simiae (AP022568.1) in order to identify a putative species-specific nucleotide sequence. A conventional PCR assay was designed to amplify a 168-bp fragment within this target, designated MST601 (M. simiae Target, 601 bp). The analytical sensitivity [Limit of Detection (LOD)] was determined using serial dilutions of genomic DNA. The pilot evaluation of the assay was assessed using 10 well-characterized clinical isolates of M. simiae. Results: The MST601-PCR assay demonstrated high analytical sensitivity, with a limit of detection of ~10 fg (≈2 genome equivalents per reaction) of M. simiae genomic DNA. No cross-reactivity among the tested species was observed with any of the 10 non-target mycobacterial species tested. The assay successfully amplified the target sequence from all 10 clinical isolates. Sequencing of the amplicons revealed ≥99% identity to reference M. simiae strains in the GenBank database, validating the assay's accuracy. Conclusion: A species-specific nucleic acid target, MST601, facilitating the rapid and accurate detection of M. simiae via conventional PCR was presented. This assay pro-vides a low-cost and accessible option for diagnostic laboratories. Molecular diagnostics, MST601, Mycobacterium simiae, Novel target, PCR, Sensitivity, Specificity 157 166 https://www.ajmb.org/En/Article.aspx?id=70652 https://www.ajmb.org/PDF/En/FullText/70652.pdf RezaKamali KakhkiMashhad Gene Azma Inc., Mashhad, Iran91889MohammadAbavisani Student Research Committee, Mashhad University of Medical Sciences, Mashhad, Iran92504KiarashGhazvini Department of Microbiology and Virology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran92505HosnaZareDepartment of Laboratory Sciences, School of Paramedical Sciences, Mashhad University of Medical Sciences, Mashhad, Iran91888NoshinHojatpanah Department of Microbiology and Virology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran92507JamalFalahi Department of Laboratory Sciences, School of Paramedical Sciences, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, Iran92508AlirezaNeshaniStudent Research Committee, Mashhad University of Medical Sciences, Mashhad, Iran31529